Monoclonal antibody analysis of lipopolysaccharide from Neisseria gonorrhoeae and Neisseria meningitidis

Apicella, M A; Bennett, K M; Hermerath, Cheryl A.; Roberts, Douglas E.

doi:10.1128/iai.34.3.751-756.1981

Cited by 83 publications

(27 citation statements)

References 11 publications

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“…LOS antigenicity was determined with a whole-cell ELISA protocol using monoclonal antibody 3F11 (Apicella et al, 1981); therefore, cells were pretreated with Clostridium perfringens Fraction V sialidase (Sigma) to expose the sialyl acceptor. Both the monoclonal antibody 3F11 and ELISA protocol, as modified from Abdillahi and Poolman (1987), were the kind gifts of P. Jones and M. Apicella (University of Iowa, Iowa City, IA).…”

Section: Elisamentioning

confidence: 99%

Sialic acid metabolism's dual function in Haemophilus influenzae

Vimr¹,

Lichtensteiger

Steenbergen

2000

Molecular Microbiology

136

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SummaryMany bacterial commensals and pathogens use the sialic acids as carbon and nitrogen sources. In Escherichia coli, the breakdown of these sugars is catalysed by gene products of the nan (N-acylneuraminate) operon; other microorganisms may use a similar catabolic strategy. Despite the known ligand and antirecognition functions of the sialic acids, the contribution of their catabolism to infection or host colonization has never been directly investigated. We addressed these questions with Haemophilus influenzae type b, which metabolizes relatively few carbohydrates, using the infant-rat infection model. The predicted H. influenzae homologue (HI0142) of the E. coli sialic acid aldolase structural gene, nanA, was subcloned and mutagenized by insertion of a kanamycin resistance cassette. Phenotypic investigation of the resulting H. influenzae aldolase mutants showed that: (i) HI0142 is essential for sialic acid degradation; (ii) the products of the open reading frames (ORFs) flanking HI0142 (HI0140, 41, 44 and 45) are likely to have the same functions as those of their counterparts in E. coli; (iii) sialylation of the lipooligosaccharide (LOS) epitope recognized by monoclonal antibody 3F11 is dependent on an environmental source of sialic acid; (iv) a nanA mutant hypersialylates its LOS sialyl acceptor, corresponding to an apparent increased fitness of the mutant in the infant-rat model; and (v) expression of the LOS sialyl acceptor is altered in cells grown without exogenous sialic acid, indicating the direct or indirect effect of sialic acid metabolism on LOS antigenicity. Taken together the data show the dual role of sialic acid catabolism in nutrition and cell surface modulation.

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Section: Elisamentioning

confidence: 99%

Sialic acid metabolism's dual function in Haemophilus influenzae

Vimr¹,

Lichtensteiger

Steenbergen

2000

Molecular Microbiology

136

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“…Despite the relatively small size of individual LOS units and the absence of repeating 0 side chains, gonococcal LOS has been demonstrated to have a complex antigenic structure with at least six antigenically distinct serotype determinants (1, 3) and two other LOS antigens which are shared by all (the common determinant) or some (the variable) strains (3). Studies with monoclonal antibody reagents have confirmed this antigenic heterogeneity (2,17). Recent studies of gonococcal surface protein antigens have focused on the phase variation that these components have been shown to undergo (5,14,28,29).…”

mentioning

confidence: 95%

Phenotypic variation in epitope expression of the Neisseria gonorrhoeae lipooligosaccharide

Apicella¹,

Shero²,

Jarvis³

et al. 1987

Infect Immun

Self Cite

121

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Gonococcal lipooligosaccharides (LOSs) are a series of antigenically complex heteropolymers. To investigate whether all members of clonally selected populations of Neisseria gonorrhoeae express antigenically similar LOS, we studied gonococcal strains 4505 and 220 with monoclonal antibodies 6B4 and 3F11 which have specificity for different oligosaccharide epitopes on the same or comigrating LOS unit(s) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fluorescent-antibody and immunoelectron microscopy studies indicated that all members of the clonally selected populations were not homogenous for the epitopes these antibodies recognized. Fluorescence-activated cell sorting studies of 3F11-coated strain 220 indicated that the density of epitope expression was a function of time of growth. The population could be separated into two broad groups corresponding to organisms staining strongly or weakly for the 3F11 epitope, and the epitope density decreased during the late-log and stationary phases of growth. Sequentially staining organisms on Formvar grids with 6B4 and 3F11, followed by staining with either 5- or 15-nm colloidal gold spheres conjugated to goat anti-mouse immunoglobulin M demonstrated the following populations of cells among organisms derived from a single clone: organisms which stained for both 6B4 and 3F11 epitopes and organisms which stained for either 6B4 epitopes alone or 3F11 epitopes alone. Immunofluorescence microscopy studies with rhodamine and fluorescein goat anti-mouse immunoglobulin M conjugates sequentially staining organisms on Formvar grids with 3F11 and 6B4 also demonstrated these three populations. Analysis of LOS preparations made over the last 5 years indicated no change in serotype antigen concentration or in sodium dodecyl sulfate-polyacrylamide gel electrophoresis migration pattern. These studies indicate that while clonally selected strains of Neisseria gonorrhoeae undergo phenotypic variation at the epitope level, the impact of this variation on the total LOS of the population has little overall effect on its antigenic or physicochemical properties.

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“…MAbs. The preparation and characterizatior 3F11, 06B4, 1-1-G, 1-1-M, and 2-1-L8 have bee previously (2,14,26,40). Mouse MAbs 2C7 an prepared by the method of Kohler and Milstein (1 fusions performed as described by Kennett (11).…”

mentioning

confidence: 99%

Antigenic and physical diversity of Neisseria gonorrhoeae lipooligosaccharides

Mandrell¹,

Schneider²,

Apicella³

et al. 1986

Infect Immun

142

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We used mouse monoclonal antibodies (MAbs) to characterize Neisseria gonorrhoeae lipooligosaccharide (LOS). LOSs that bound two or more MAbs in a solid-phase radioimmunoassay usually bound them to different LOS components, as separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); strains with multiple LOS components on SDS-PAGE usually bound more than one MAb. However, the LOS of some strains bound the same MAb to two LOS components with different relative molecular weights, and some individual LOS components bound more than one MAb. LOSs from different strains bound different amounts of the same MAb at saturation, reflecting differences in the quantitative expression of individual LOS components. Not all components recognized by MAbs were stained by silver after periodate oxidation. Treatment with NaOH variously affected epitopes defined by different MAbs. MAb 3F11 completely inhibited and MAb 2-1-L8 partially inhibited the binding of 125I-labeled 06B4 MAb to WR220 LOS and WR220 outer membranes in competitive binding studies. Other MAbs did not compete with the binding of '2'I-labeled 06B4 to either antigen. We conclude that a strain of N. gonorrhoeae elaborates multiple LOSs that can be separated by SDS-PAGE and that are antigenically distinct. Epitope expression within these glycolipids is complex. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separates the principle outer membrane glycolipids, or lipooligosaccharides (LOSs), of Neisseria gonorrhoeae into multiple components with Mrs between 3,000 and 7,000, as determined with isogenic Salmonella rough mutant LOS standards (25, 26). High Mr components bearing polysaccharides composed of increasing numbers of oligosaccharide repeating units that are produced by smooth strains of members of the family Enterobacteriaceae (lipopolysaccharides [LPSs]) (7) are absent from gonococcal LOS. Gonococcal LOSs that migrate in the same molecular weight range as the bands observed with smooth enterobacterial LPS are LOS aggregates that diminish when treated with NaOH or when urea is included in the gel (16). For these reasons the term LOS has been used (8, 14, 24, 26, 33, 36) to distinguish the endotoxin moiety of Neisseria from the generally larger endotoxin molecules of members of the family Enterobacteriaceae. Antigen expression within gonococcal LOS is heterogeneous, reflecting the heterogeneity of molecular size (26). The complex antigenic structures of their LOSs have made serological classification of gonococci difficult and thwarted efforts to understand human immunity (1, 3, 5, 9, 16-18, 23, 24). Monoclonal antibodies potentially could provide ideal tools for defining the physical basis of antigen expression within complex glycolipids (6). Mouse monoclonal antibodies (MAbs) that bind to gonococcal LOSs have been described previously (2, 10, 14, 26). An immunoglobulin M (IgM) MAb, 3F11, recognizes a * Corresponding author. t This is report no. 2 from the Centre for Immunochemistry of the

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Monoclonal antibody analysis of lipopolysaccharide from Neisseria gonorrhoeae and Neisseria meningitidis

Cited by 83 publications

References 11 publications

Sialic acid metabolism's dual function in Haemophilus influenzae

Sialic acid metabolism's dual function in Haemophilus influenzae

Phenotypic variation in epitope expression of the Neisseria gonorrhoeae lipooligosaccharide

Antigenic and physical diversity of Neisseria gonorrhoeae lipooligosaccharides

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